Morphological and molecular genetic characterization of Kudoa konishiae n. sp. (Myxosporea: Multivalvulida) in the muscle of Japanese Spanish mackerel (Scomberomorus niphonius).

The Japanese Spanish mackerel (Scomberomorus niphonius; Perciformes: Scombridae) is widely distributed in the continental shelves of the northwestern Pacific Ocean around Japan, Sea of Japan, and East China Sea. In the present study, small, spindle-shaped myxosporean plasmodia (0.15-0.53 mm by 0.04-0.13 mm) were frequently encountered in the myofiber of trunk muscles of two Japanese Spanish mackerels; one fished in the Sea of Japan off western Japan and the other in the northwestern Pacific Ocean off southern Japan in the autumn of 2016. Isolated myxospores of Kudoa konishiae n. sp. (Myxosporea: Multivalvulida) from these two fish were stellate with six equal shell valves and polar capsules, 8.1-9.7 μm in width, 7.1-8.8 μm in thickness, and 7.1-8.8 μm in length. The polar capsules were teardrop-shaped, 2.7-4.7 μm by 1.2-2.5 μm. The lateral view of spores revealed a drawstring-pouch shape. The nucleotide sequences of the 18S and 28S ribosomal RNA gene (rDNA) were distinct from any recorded species. Phylogenetic trees demonstrated a close relationship of the present new species with Kudoa spp. with stellate spores with five or more shell valves/polar capsules, recorded in scombrid fishes. To clarify the phylogenetic relationships between three closely related species, i.e., Kudoa konishiae n. sp., Kudoa hexapunctata, and Kudoa neothunni, three mitochondrial DNA genes (cytochrome c oxidase subunit 1 gene (cox-1) and the small and large subunits of the ribosomal RNA gene (rns-rnl)) of two isolates of the new species, six isolates of K. hexapunctata, and 13 isolates of K. neothunni were sequenced. The interspecific and intraspecific variations of the newly obtained cox-1 and rns-rnl nucleotide sequences of K. hexapunctata, K. neothunni, and K. konishiae n. sp. were clarified for the first time.